Animal integration vector and methods for its use

Abstract

An animal integration vector and methods for its use in the insertion of an exogenous nucleic acid into the genome of a whole animal are provided. The vectors of the subject invention include a pair of transposase recognized insertion sequences, e.g., P element transposase recognized insertion sequences (such as P element derived 31 base pair inverted repeats), flanking at least one transcriptionally active gene that is located sufficienty proximal to one of the transposase recognized insertion sequences, e.g., P-feet, to provide for the desired genome integration. In practicing the subject methods, a vector as described above carrying an exogenous or endogenous nucleic acid is introduced into a target animal under conditions sufficient for entry into the animal and its cells resulting in transposition of the exogenous nucleic acid from the vector into the animal genome, i.e., in the genome of at least a portion of the cells making up the animal. The subject methods find use in a variety of transformation applications, including research, polypeptide synthesis and therapeutic applications.

1 - 10 . (Canceled).  
     
     
         11 . A method of inserting an exogenous nucleic acid into the genome of a non-Drosophilidae animal, said method comprising: 
 introducing into said animal a transposase recognized insertion sequence vector comprising said exogenous nucleic acid under conditions sufficient for transposition to occur so that said exogenous nucleic acid is inserted into said genome.    
     
     
         12 . A method of inserting an exogenous nucleic acid into the genome of a non-Drosophilidae animal, said method comprising: 
 introducing into said animal a vector according to claim  1  under conditions sufficient for transposition to occur so that said exogenous nucleic acid is inserted into said genome.    
     
     
         13 . The method according to  claim 12 , wherein said vector comprises a transposase domain.  
     
     
         14 . The method according to  claim 12 , wherein said method further comprises introducing a second vector comprising a transposase domain into said animal.  
     
     
         15 . The method according to  claim 12 , wherein said exogenous nucleic acid ranges in length from about 50 to 150,000 bp.  
     
     
         16 . The method according to  claim 12 , wherein said target animal is a vertebrate.  
     
     
         17 . The method according to  claim 12 , wherein said vertebrate animal is a mammalian animal.  
     
     
         18 . The method according to  claim 12 , wherein said mammalian animal is a rodent.  
     
     
         19 . A kit for use in inserting an exogenous nucleic acid into a target cell, said kit comprising: 
 a P-element vector comprising a pair of P-element transposase recognized insertion sequences flanking at least one transcriptionally active gene in proximity to at least one of the P-element transposase recognized isertion sequences.    
     
     
         20 . The kit according to  claim 19 , wherein said transcriptionally active gene comprises a coding sequence that is expressed under intracellular conditions.  
     
     
         21 . The kit according to  claim 19 , wherein said vector further comprises at least one endonuclease cleavage site positioned between said transposase recognized insertion sequences.  
     
     
         22 . The kit according to  claim 21 , wherein said endonuclease cleavage site is present in a polylinker.  
     
     
         23 - 26 . (Canceled)  
     
     
         27 . A non-Drosophilidae animal or cells derived from said animal that has P-element transposase recognized insertion sequences integrated into the genome.  
     
     
         28 . The animal or cells according to  claim 27 , wherein said animal is a vertebrate or said cells are vertebrate cells.  
     
     
         29 . The animal or cells according to  claim 28 , wherein said animal is a mammal or said cells are mammalian cells.  
     
     
         30 . The animal or cells according to  claim 29 , wherein said animal is a rodent or said cells are rodent cells.  
     
     
         31 . A non-Drosophilidae animal or cells derived from said animal that have P element transposase recognized 31 bp insertion sequences integrated into the genome.  
     
     
         32 . The animal or cells according to  claim 31 , wherein said animal is a vertebrate or said cells are vertebrate cells.  
     
     
         33 . The animal or cells according to  claim 32 , wherein said animal is a mammal or said cells are mammalian cells.  
     
     
         34 . The animal or cells according to  claim 33 , wherein said animal is a rodent or said cells are rodent cells.